Inhibitors of macrophage migration inhibitory factor

ABSTRACT

A method for reducing macrophage migration inhibitory factor (MIF or MMIF) cytokine or its biological activity, including the step of administering an isothiocyanate functional surfactant to a patient having a disease or condition wherein MIF cytokine or its biological activity is implicated in the disease or condition. In one embodiment, the protonated form of the isothiocyanate functional surfactant is represented by the following chemical structure:

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.16/025,579, entitled “INHIBITORS OF MACROPHAGE MIGRATION INHIBITORYFACTOR” filed Jul. 2, 2018, now U.S. Pat. No. 10,434,081, which is acontinuation-in-part of U.S. application Ser. No. 15/838,444, entitled“METHOD FOR TREATING BLADDER CANCER” filed Dec. 12, 2017, now U.S. Pat.No. 10,111,852, which is a continuation of U.S. application Ser. No.15/423,869, entitled “METHOD FOR TREATING BLADDER CANCER” filed Feb. 3,2017, now U.S. Pat. No. 9,839,621, which is a continuation-in-part ofU.S. application Ser. No. 14/867,626, entitled “METHOD FOR TREATING SKINCANCER,” filed Sep. 28, 2015, now U.S. Pat. No. 9,642,827, which is acontinuation of U.S. application Ser. No. 14/867,585, entitled “METHODFOR TREATING SKIN CANCER,” filed Sep. 28, 2015, now U.S. Pat. No.9,636,320, which is a continuation of U.S. application Ser. No.14/519,510, entitled “METHOD FOR TREATING SKIN CANCER,” filed Oct. 21,2014, now U.S. Pat. No. 9,504,667, which is a continuation of U.S.application Ser. No. 13/952,236, entitled “METHOD FOR TREATING SKINCANCER,” filed Jul. 26, 2013, now U.S. Pat. No. 8,865,772, which claimsthe benefit of U.S. Provisional Application Ser. No. 61/676,093,entitled “METHOD FOR TREATING SKIN CANCER,” filed Jul. 26, 2012—whichare hereby incorporated herein by reference in their entirety, includingall references cited therein.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

Not applicable.

REFERENCE TO A SEQUENCE LISTING

Not applicable.

BACKGROUND OF THE INVENTION 1. Field of the Invention

The present invention relates in general to a method for reducingmacrophage migration inhibitory factor (MIF or MMIF) cytokine (alsoknown as glycosylation-inhibiting factor (GIF)) or its biologicalactivity, and, more particularly, to a method for treating and/orpreventing a disease or condition, wherein MIF cytokine or itsbiological activity is implicated in the disease or condition, with anisothiocyanate functional surfactant (ISF).

2. Background Art

The protein MIF is a cytokine released by many cell types includingT-lymphocytes and macrophages. MIF proteins have been identified inseveral species including mammals and are generally 12-13 kDa in size.MIF levels are believed to increase during physiological stress orsystemic inflammatory conditions. MIF plays an important role in septicshock and delayed-type hypersensitivity reactions, possibly due to itsability to act as an endogenous regulator of glucocorticoid actionwithin the immune system. Deletion of the MIF gene orimmunoneutralization of MIF protects against septic shock.

Elevated levels of MIF have been observed in a number of disease statesincluding cardiovascular disease, diabetes, sepsis and many types ofcancer. Furthermore, genetic ablation of MIF has been shown to attenuatevarious disease states in murine models. While details surrounding themechanisms of MIF action are still unclear, the clinical significance ofMIF expression is such that targeted approaches to modulate thebiological activities of MIF are currently in development—see thecomplete file wrapper history of International Publication Number WO2010/140902 A1 Entitled “Inhibitors Of Macrophage Migration InhibitoryFactor,” which is hereby incorporated herein by reference in itsentirety, including all references cited therein.

MIF is also known to have a proinflammatory role in arthritis andglomerulonephritis. Inhibition of proinflammatory cytokine activityusing monoclonal antibodies has been shown to improve disease outcomesin mouse models of rheumatoid arthritis, sepsis, cardiovascular disease,and inflammatory bowel disease. Additional biological activities for MIFinclude tumor invasion, metastasis and angiogenesis, insulin release,cell growth and apoptosis, regulation of T-cell and macrophageactivation and IgE synthesis. MIF also acts as a tautomerase. While thebiological significance of this activity is still under investigation,the tautomerase site appears to be important for regulatingprotein-protein interactions that mediate MIF activity.

MIF is known to be involved in numerous pathological events, andinhibition of MIF may have several therapeutic effects. Accordingly, itis an object of the present invention to provide new and usefulcompounds that serve as MIF inhibitors and/or modulators.

SUMMARY OF THE INVENTION

In one embodiment, the present invention is directed to a method forreducing macrophage migration inhibitory factor cytokine or itsbiological activity comprising the step of: administering anisothiocyanate functional surfactant to a patient having a disease orcondition wherein MIF cytokine or its biological activity is implicatedin the disease or condition.

In a preferred embodiment of the present invention, the isothiocyanatefunctional surfactant comprises at least one isothiocyanate functionalgroup associated with an aliphatic and/or aromatic carbon atom of theisothiocyanate functional surfactant.

In another preferred embodiment of the present invention, theisothiocyanate functional surfactant comprises a lysine derivative,wherein the lysine derivative comprises an α-nitrogen and a ε-nitrogen,and wherein an alkyl and/or alkanoyl substituent comprising at leastapproximately 8 carbon atoms is associated with the α-nitrogen, andfurther wherein at least one isothiocyanate functional group isassociated with the ε-nitrogen.

In yet another preferred embodiment of the present invention, theisothiocyanate functional surfactant is represented by the followingchemical structure:

wherein the protonated form of the surfactant comprises a non-polarmoiety (NP) and a polar moiety (P), and wherein at least oneisothiocyanate functional group (NCS) is associated with the polarand/or non-polar moiety.

In another aspect of the present invention, the protonated form of theisothiocyanate functional surfactant is represented by the followingchemical structure:

wherein R₁ comprises an alkyl, cycloalkyl, polycycloalkyl,heterocycloalkyl, aryl, alkaryl, aralkyl, alkoxy, alkanoyl, aroyl,alkenyl, alkynyl and/or cyano group containing approximately 1 toapproximately 25 carbon atom(s), wherein the carbon atom(s) may be alinking group to, or part of, a halogen, a N, O, and/or S containingmoiety, and/or one or more functional groups comprising alcohols,esters, ammonium salts, phosphonium salts, and combinations thereof; alinkage to a dimer; a linkage to an oligomer; and/or a linkage to apolymer; wherein R₂ comprises NCS; and wherein R₃-R₅ are the same ordifferent and comprise H; OH; an alkyl, cycloalkyl, polycycloalkyl,heterocycloalkyl, aryl, alkaryl, aralkyl, alkoxy, alkanoyl, aroyl,alkenyl, alkynyl and/or cyano group containing approximately 1 toapproximately 25 carbon atom(s), wherein the carbon atom(s) may be alinking group to, or part of, a halogen, a N, O, and/or S containingmoiety, and/or one or more functional groups comprising alcohols,esters, ammonium salts, phosphonium salts, and combinations thereof; alinkage to a dimer; a linkage to an oligomer; and/or a linkage to apolymer with the proviso that at least one of R₃-R₅ comprise an alkyl,cycloalkyl, polycycloalkyl, heterocycloalkyl, aryl, alkaryl, aralkyl,alkoxy, alkanoyl, aroyl, alkenyl, alkynyl and/or cyano group containingapproximately 8 to approximately 25 carbon atom(s).

In a preferred embodiment of the present invention, the protonated formof the isothiocyanate functional surfactant is represented by thefollowing chemical structure:

wherein R₁ is selected from the group consisting of an alkyl groupcontaining 1 to 25 carbon atom(s); wherein R₂ is selected from the groupconsisting of NCS; and wherein R₃-R₅ are each independently selectedfrom the group consisting of H; OH; and an alkyl, and alkanoyl groupcontaining 1 to 25 carbon atom(s) with the proviso that at least one ofR₃-R₅ is selected from the group consisting of an alkyl, and alkanoyl,group containing 8 to 25 carbon atoms.

In another preferred embodiment of the present invention, the protonatedform of the isothiocyanate functional surfactant is represented by thefollowing chemical structure:

wherein X comprises an integer ranging from approximately 1 toapproximately 25, and wherein Y comprises an integer ranging fromapproximately 6 to approximately 25.

In yet another preferred embodiment of the present invention, theprotonated form of the isothiocyanate functional surfactant isrepresented by the following chemical structure:

In a preferred embodiment of the present invention, the protonated formof the isothiocyanate functional surfactant is represented by at leastone of the following chemical structures:

In another preferred embodiment of the present invention, theisothiocyanate functional surfactant is represented by the followingchemical structure:

wherein R₁ comprises an alkyl, cycloalkyl, polycycloalkyl,heterocycloalkyl, aryl, alkaryl, aralkyl, alkoxy, alkanoyl, aroyl,alkenyl, alkynyl and/or cyano group containing approximately 1 toapproximately 25 carbon atom(s), wherein the carbon atom(s) may be alinking group to, or part of, a halogen, a N, O, and/or S containingmoiety, and/or one or more functional groups comprising alcohols,esters, ammonium salts, phosphonium salts, and combinations thereof; alinkage to a dimer; a linkage to an oligomer; and/or a linkage to apolymer; wherein R₂ comprises NCS; wherein R₃-R₅ are the same ordifferent and comprise H; OH; an alkyl, cycloalkyl, polycycloalkyl,heterocycloalkyl, aryl, alkaryl, aralkyl, alkoxy, alkanoyl, aroyl,alkenyl, alkynyl and/or cyano group containing approximately 1 toapproximately 25 carbon atom(s), wherein the carbon atom(s) may be alinking group to, or part of, a halogen, a N, O, and/or S containingmoiety, and/or one or more functional groups comprising alcohols,esters, ammonium salts, phosphonium salts, and combinations thereof; alinkage to a dimer; a linkage to an oligomer; and/or a linkage to apolymer with the proviso that at least one of R₃-R₅ comprise an alkyl,cycloalkyl, polycycloalkyl, heterocycloalkyl, aryl, alkaryl, aralkyl,alkoxy, alkanoyl, aroyl, alkenyl, alkynyl and/or cyano group containingapproximately 8 to approximately 25 carbon atom(s), wherein X comprisesa counter cation such as, but not limited to, alkali metals, alkalineearth metals, transition metals, s-block metals, d-block metals, p-blockmetals, NZ₄ ⁺, wherein Z comprises, H, R₆, OR₆, and wherein R₆ comprisesan alkyl, cycloalkyl, polycycloalkyl, heterocycloalkyl, aryl, alkaryl,aralkyl, alkoxy, alkanoyl, aroyl, alkenyl, alkynyl and/or cyano groupcontaining approximately 1 to approximately 25 carbon atom(s), whereinthe carbon atom(s) may be a linking group to, or part of, a halogen, aN, O, and/or S containing moiety, and/or one or more functional groupscomprising alcohols, esters, ammonium salts, phosphonium salts, andcombinations thereof; a linkage to a dimer; a linkage to an oligomer;and/or a linkage to a polymer.

In yet another preferred embodiment of the present invention, theisothiocyanate functional surfactant is represented by the followingchemical structure:

wherein R₁ is selected from the group consisting of an alkyl groupcontaining 1 to 25 carbon atom(s); wherein R₂ is selected from the groupconsisting of NCS; and wherein R₃-R₅ are each independently selectedfrom the group consisting of H; OH; and an alkyl, and alkanoyl groupcontaining 1 to 25 carbon atom(s) with the proviso that at least one ofR₃-R₅ is selected from the group consisting of an alkyl, and alkanoyl,group containing 8 to 25 carbon atoms; and wherein X comprises a countercation.

In another aspect of the present invention, the isothiocyanatefunctional surfactant is administered to the patient at least one oforally, intravenously, intramuscularly, intrathecally, cutaneously,subcutaneously, transdermally, sublingually, buccally, rectally,vaginally, ocularly, otically, and nasally.

In a preferred embodiment of the present invention, the isothiocyanatefunctional surfactant further comprises one or more pharmaceutical,biological or molecular biological active agents.

DETAILED DESCRIPTION OF THE INVENTION

While this invention is susceptible of embodiment in many differentforms, there is shown in the drawings and/or described herein in detailseveral specific embodiments with the understanding that the presentdisclosure is to be considered as an exemplification of the principlesof the invention and is not intended to limit the invention to theembodiments illustrated.

In accordance with the present invention, methods for reducing and/ormodulating macrophage migration inhibitory factor cytokine or itsbiological activity are provided herein. In particular, these methodsinclude treating a plurality of types of diseases and/or conditions,such as, but not limited to, rheumatoid arthritis, sepsis,cardiovascular disease, diabetes, inflammatory bowel disease, andcancer—just to name a few.

In one embodiment, the present invention is directed to method foreliminating and/or reducing macrophage migration inhibitory factorcytokine or its biological activity by administering an isothiocyanatefunctional surfactant to a patient having a disease or condition whereinMIF cytokine or its biological activity is implicated in the disease orcondition. Preferably, the isothiocyanate functional surfactantcomprises one or more isothiocyanate functional groups associated withan aliphatic and/or aromatic carbon atom of the isothiocyanatefunctional surfactant. It will be understood that isothiocyanatefunctional surfactants, regardless of their ordinary meaning, aredefined herein as a surfactant having an isothiocyanate functional groupassociated therewith. It will be yet further understood that the term“associated” as used herein in chemical context, regardless of itsordinary meaning, is defined herein as attached, a covalent bond, apolar covalent bond, an ionic bond, a hydrogen bond, van der Waalsforces, electrostatic interaction, directly and/or indirectly linked,etcetera.

The term surfactant derives from contraction of the termssurface-active-agent and is defined herein as a molecule and/or group ofmolecules which are able to modify the interfacial properties of theliquids (aqueous and non-aqueous) in which they are present. Thesurfactant properties of these molecules reside in their amphiphiliccharacter which stems from the fact that each surfactant molecule hasboth a hydrophilic moiety and a hydrophobic (or lipophilic) moiety, andthat the extent of each of these moieties is balanced so that atconcentrations at or below the critical micelle concentration (i.e.,CMC) they generally concentrate at the air-liquid interface andmaterially decrease the interfacial tension. For example, sodium saltsof saturated carboxylic acids are extremely soluble in water up to C8length and are thus not true surfactants. They become less soluble inwater from C9 up to C18 length, the domain of effective surfactants forthis class of compounds. The carboxylic acids (fatty acids) can beeither saturated or unsaturated starting from C16 chain lengths.

Without being bound by any one particular theory, it is believed thatthe isothiocyanate functional surfactants disclosed herein facilitatetreatment of numerous diseases and conditions by eliminating,modulating, and/or reducing macrophage migration inhibitory factorcytokine or its biological activity. It is also believed that theisothiocyanate functional surfactants disclosed herein facilitateelevating phase II enzymes (e.g., HAD(P)H quinine oxidoreductase) whichare believed to, among other things regulate inflammatory responseswithin the body, as well as detoxify carcinogens and/or activatedcarcinogens.

In accordance with the present invention, the isothiocyanate functionalsurfactants may be used as an administered leave-on/leave-in product inwhich one or more surfactants remain on/in the body and are notimmediately and/or ever removed from the body. Alternatively, theisothiocyanate functional surfactants of the present invention may beused in an administer and remove fashion. For either case, it ispreferred that the isothiocyanate functional surfactants be generallymild to human body (e.g., non-irritating or low-irritating). Inparticular, anionic N-alkanoyl surfactants derived from amino acids areespecially preferred because, while not completely predictable, theyhave a tendency to be mild. The methods of preparation detailed in thisinvention employ, but are not limited to, amino acids that possess atleast two amine functionalities, at least one of which is converted toan N-alkanoyl functionality, and at least one of which is converted intoisothiocyanate functionality. The amino acids include, but are notlimited to, the α-amino acids lysine, ornithine, 2,4-diaminobutanoicacid, 2,3-diaminopropionic acid, 2,7-diaminoheptanoic acid, and2,8-diaminooctanoic acid. Additionally, amino acids other than α-aminoacids may be employed, such as β-amino acids, etcetera. It will beunderstood that amino acid derived surfactants are preferred due totheir mild nature, but any one of a number of other surfactants arelikewise contemplated for use in accordance with the present invention.

Methods for preparing isothiocyanate functional surfactants and/or theirprecursors can involve, but are not limited to, conversion of an aminefunctionality to an isothiocyanate functionality. The methods ofconversion of amine functionalities to isothiocyanate functionalitiesinclude, but are not limited to: (1) reaction with carbon disulfide toyield an intermediate dithiocarbamate, followed by reaction withethylchloroformate or its functional equivalent such asbis(trichloromethyl)-carbonate, trichloromethyl chloroformate, orphosgene; (2) reaction with thiophosgene; (3) reaction with1,1′-thiocarbonyldiimidizole; (4) reaction with phenylthiochloroformate;(5) reaction with ammonium or alkali metal thiocyanate to prepare anintermediate thiourea followed by cleaving to the isothiocyanate viaheating; and (6) reaction with an isothiocyanato acyl halide[SCN—(CH₂)_(n)—CO—Cl]. The resulting isothiocyanate functionalsurfactant, depending on the method of preparation, can be isolated as apure material or as a mixture with other surfactants. The resultingisothiocyanate functional surfactant, depending on the method ofpreparation, can be isolated and used directly in nonionic form, anionicform, cationic form, zwitterionic (amphoteric) form, and/or in a neutralsurfactant-precursor form in combination with a base such as sodiumhydroxide or triethanol amine if the neutral surfactant-precursor formpossesses a protonated carboxylic acid group such that reaction(deprotonation) with the base converts the neutral surfactant-precursorform to an anionic surfactant, or in neutral surfactant-precursor formin combination with an acid if the neutral surfactant-precursor formpossess amine functionality such that reaction (protonation) with theacid converts the neutral surfactant-precursor form to a cationicsurfactant.

In accordance with the present invention, the step of administeringcomprises, but is not limited to, systemic administration, localinjection, regional injection, spraying, dripping, dabbing, rubbing,blotting, dipping, and any combination thereof.

In one preferred embodiment of the present invention, the isothiocyanatefunctional surfactant is removed from body and/or affected area after aperiod of time. Such a period comprises, but is not limited to, seconds(e.g., 1 second, 2 seconds, 5 seconds, 10 seconds, 15 seconds, 20seconds, 30 seconds, 45 seconds, and 60 seconds), minutes (e.g., 1minute, 2 minutes, 5 minutes, 10 minutes, 15 minutes, 20 minutes, 30minutes, 45 minutes, and 60 minutes), hours (e.g., 1 hour, 2 hours, 4hours, 5 hours, 8 hours, 10 hours, 15 hours, 24 hours, 36 hours, 48hours, and 60 hours), days (e.g., 1 day, 2 days, 3 days, 4 days, 5 days,6 days, 7 days, 10 days, 14 days, 21 days, 30 days), etcetera. It willbe understood that the step of removing preferably occurs via purging,rinsing, wiping, and/or extracting—just to name a few.

Depending upon the subject and/or the severity of the condition and/ordisease, multiple administrations may be necessary. As such, the stepsof administering and/or removing the isothiocyanate functionalsurfactant may be repeated one or a plurality of times.

The present invention is also directed to a method for reducingmacrophage migration inhibitory factor cytokine or its biologicalactivity comprising the steps of associating (using any known medicaltechnique) a lysine derivative to a patient having a disease orcondition wherein MIF cytokine or its biological activity is implicatedin the disease or condition, wherein the lysine derivative comprises anα-nitrogen and a ε-nitrogen. Preferably, an alkyl substituent comprisingat least approximately 8 carbon atoms is associated with the α-nitrogen.Preferably, at least one isothiocyanate functional group is associatedwith the ε-nitrogen.

The present invention is further directed to a method for reducingmacrophage migration inhibitory factor cytokine or its biologicalactivity comprising the step of: administering a surfactant to a patienthaving a disease or condition wherein MIF cytokine or its biologicalactivity is implicated in the disease or condition, wherein thesurfactant is represented by the following chemical structure:

and wherein the surfactant comprises a non-polar moiety (NP) and a polarmoiety (P), and wherein at least one isothiocyanate functional group(NCS) is associated with the polar and/or non-polar moiety.

The present invention is yet further directed to a method for reducingmacrophage migration inhibitory factor cytokine or its biologicalactivity comprising the step of: administering a surfactant to a patienthaving a disease or condition wherein MIF cytokine or its biologicalactivity is implicated in the disease or condition, wherein theprotonated form of the surfactant is represented by the followingchemical structure:

wherein R₁ comprises an alkyl, cycloalkyl, polycycloalkyl,heterocycloalkyl, aryl, alkaryl, aralkyl, alkoxy, alkanoyl, aroyl,alkenyl, alkynyl and/or cyano group containing approximately 1 toapproximately 25 carbon atom(s), wherein the carbon atom(s) may be alinking group to, or part of, a halogen, a N, O, and/or S containingmoiety, and/or one or more functional groups comprising alcohols,esters, ammonium salts, phosphonium salts, and combinations thereof; alinkage to a dimer; a linkage to an oligomer; and/or a linkage to apolymer; wherein R₂ comprises NCS; and wherein R₃-R₅ are the same ordifferent and comprise H; OH; an alkyl, cycloalkyl, polycycloalkyl,heterocycloalkyl, aryl, alkaryl, aralkyl, alkoxy, alkanoyl, aroyl,alkenyl, alkynyl and/or cyano group containing approximately 1 toapproximately 25 carbon atom(s), wherein the carbon atom(s) may be alinking group to, or part of, a halogen, a N, O, and/or S containingmoiety, and/or one or more functional groups comprising alcohols,esters, ammonium salts, phosphonium salts, and combinations thereof; alinkage to a dimer; a linkage to an oligomer; and/or a linkage to apolymer with the proviso that at least one of R₃-R₅ comprise an alkyl,cycloalkyl, polycycloalkyl, heterocycloalkyl, aryl, alkaryl, aralkyl,alkoxy, alkanoyl, aroyl, alkenyl, alkynyl and/or cyano group containingapproximately 8 to approximately 25 carbon atom(s).

In this embodiment, the surfactant is preferably represented by thefollowing chemical structure:

wherein X comprises an integer ranging from approximately 1 toapproximately 25, and wherein Y comprises an integer ranging fromapproximately 6 to approximately 25.

More preferably, the surfactant is represented by the following chemicalstructure:

In a preferred embodiment of the present invention, the protonated formof the isothiocyanate functional surfactant is represented by at leastone of the following chemical structures:

In another embodiment, the present invention is directed to a methodreducing macrophage migration inhibitory factor cytokine or itsbiological activity comprising the step of: administering a surfactantto a patient having a disease or condition wherein MIF cytokine or itsbiological activity is implicated in the disease or condition, whereinthe protonated form of the surfactant is represented by the followingchemical structure:

wherein R₁ comprises an alkyl, cycloalkyl, polycycloalkyl,heterocycloalkyl, aryl, alkaryl, aralkyl, alkoxy, alkanoyl, aroyl,alkenyl, alkynyl and/or cyano group containing approximately 1 toapproximately 25 carbon atom(s), wherein the carbon atom(s) may be alinking group to, or part of, a halogen, a N, O, and/or S containingmoiety, and/or one or more functional groups comprising alcohols,esters, ammonium salts, phosphonium salts, and combinations thereof; alinkage to a dimer; a linkage to an oligomer; and/or a linkage to apolymer; wherein R₂ comprises NCS; wherein R₃-R₅ are the same ordifferent and comprise H; OH; an alkyl, cycloalkyl, polycycloalkyl,heterocycloalkyl, aryl, alkaryl, aralkyl, alkoxy, alkanoyl, aroyl,alkenyl, alkynyl and/or cyano group containing approximately 1 toapproximately 25 carbon atom(s), wherein the carbon atom(s) may be alinking group to, or part of, a halogen, a N, O, and/or S containingmoiety, and/or one or more functional groups comprising alcohols,esters, ammonium salts, phosphonium salts, and combinations thereof; alinkage to a dimer; a linkage to an oligomer; and/or a linkage to apolymer with the proviso that at least one of R₃-R₅ comprise an alkyl,cycloalkyl, polycycloalkyl, heterocycloalkyl, aryl, alkaryl, aralkyl,alkoxy, alkanoyl, aroyl, alkenyl, alkynyl and/or cyano group containingapproximately 8 to approximately 25 carbon atom(s), wherein X comprisesa counter cation such as, but not limited to, alkali metals, alkalineearth metals, transition metals, s-block metals, d-block metals, p-blockmetals, NZ₄ ⁺, wherein Z comprises, H, R₆, and/or OR₆, and wherein R₆comprises an alkyl, cycloalkyl, polycycloalkyl, heterocycloalkyl, aryl,alkaryl, aralkyl, alkoxy, alkanoyl, aroyl, alkenyl, alkynyl and/or cyanogroup containing approximately 1 to approximately 25 carbon atom(s),wherein the carbon atom(s) may be a linking group to, or part of, ahalogen, a N, O, and/or S containing moiety, and/or one or morefunctional groups comprising alcohols, esters, ammonium salts,phosphonium salts, and combinations thereof; a linkage to a dimer; alinkage to an oligomer; and/or a linkage to a polymer.

In accordance with the present invention, the isothiocyanate functionalsurfactant may also be associated with one or more additionalsurfactants, wherein the additional surfactants are selected from atleast one of the group comprising a non-ionic surfactant, an anionicsurfactant, a cationic surfactant, a zwitterionic surfactant, andcombinations thereof.

Non-limiting examples of preferred anionic surfactants include taurates;isethionates; alkyl and alkyl ether sulfates; succinamates; alkylsulfonates, alkylaryl sulfonates; olefin sulfonates; alkoxy alkanesulfonates; sodium and potassium salts of fatty acids derived fromnatural plant or animal sources or synthetically prepared; sodium,potassium, ammonium, and alkylated ammonium salts of alkylated andacylated amino acids and peptides; alkylated sulfoacetates; alkylatedsulfosuccinates; acylglyceride sulfonates, alkoxyether sulfonates;phosphoric acid esters; phospholipids; and combinations thereof.Specific anionic surfactants contemplated for use include, but are by nomeans limited to, ammonium cocoyl isethionate, sodium cocoylisethionate, sodium lauroyl isethionate, sodium stearoyl isethionate,sodium lauroyl sarcosinate, sodium cocoyl sarcosinate, sodium laurylsarcosinate, disodium laureth sulfosuccinate, sodium laurylsulfoacetate, sodium cocoyl glutamate, TEA-cocoyl glutamate, TEA cocoylalaninate, sodium cocoyl taurate, potassium cetyl phosphate.

Non-limiting examples of preferred cationic surfactants includealkylated quaternary ammonium salts R₄NX; alkylated amino-amides(RCONH—(CH₂)_(n))NR₃X; alkylimidazolines; alkoxylated amines; andcombinations thereof. Specific examples of anionic surfactantscontemplated for use include, but are by no means limited to, cetylammonium chloride, cetyl ammonium bromide, lauryl ammonium chloride,lauryl ammonium bromide, stearyl ammonium chloride, stearyl ammoniumbromide, cetyl dimethyl ammonium chloride, cetyl dimethyl ammoniumbromide, lauryl dimethyl ammonium chloride, lauryl dimethyl ammoniumbromide, stearyl dimethyl ammonium chloride, stearyl dimethyl ammoniumbromide, cetyl trimethyl ammonium chloride, cetyl trimethyl ammoniumbromide, lauryl trimethyl ammonium chloride, lauryl trimethyl ammoniumbromide, stearyl trimethyl ammonium chloride, stearyl trimethyl ammoniumbromide, lauryl dimethyl ammonium chloride, stearyl dimethyl cetylditallow dimethyl ammonium chloride, dicetyl ammonium chloride, dilaurylammonium chloride, dilauryl ammonium bromide, distearyl ammoniumchloride, distearyl ammonium bromide, dicetyl methyl ammonium chloride,dicetyl methyl ammonium bromide, dilauryl methyl ammonium chloride,distearyl methyl ammonium chloride, distearyl methyl ammonium bromide,ditallow dimethyl ammonium chloride, ditallow dimethyl ammonium sulfate,di(hydrogenated tallow) dimethyl ammonium chloride, di(hydrogenatedtallow) dimethyl ammonium acetate, ditallow dipropyl ammonium phosphate,ditallow dimethyl ammonium nitrate, di(coconutalkyl)dimethyl ammoniumchloride, di(coconutalkyl)dimethyl ammonium bromide, tallow ammoniumchloride, coconut ammonium chloride, stearamidopropyl PG-imoniumchloride phosphate, stearamidopropyl ethyldimonium ethosulfate,stearimidopropyldimethyl (myristyl acetate) ammonium chloride,stearamidopropyl dimethyl cetearyl ammonium tosylate, stearamidopropyldimethyl ammonium chloride, stearamidopropyl dimethyl ammonium lactate,ditallowyl oxyethyl dimethyl ammonium chloride, behenamidopropyl PGdimonium chloride, dilauryl dimethyl ammonium chloride, distearlydimethyl ammonium chloride, dimyristyl dimethyl ammonium chloride,dipalmityl dimethyl ammonium chloride, distearyl dimethyl ammoniumchloride, stearamidoproyl PG-dimonium chloride phosphate,stearamidopropyl ethyldiammonium ethosulfate, stearamidopropyl dimethyl(myristyl acetate) ammonium chloride, stearimidopropyl dimethyl cetarylammonium tosylate, stearamido propyl dimethyl ammonium chloride,stearamidopropyl dimethyl ammonium lactate.

Non-limiting examples of preferred non-ionic surfactants includealcohols, alkanolamides, amine oxides, esters (including glycerides,ethoxylated glycerides, polyglyceryl esters, sorbitan esters,carbohydrate esters, ethoxylated carboxylic acids, phosphoric acidtriesters), ethers (including ethoxylated alcohols, alkyl glucosides,ethoxylated polypropylene oxide ethers, alkylated polyethylene oxides,alkylated polypropylene oxides, alkylated PEG/PPO copolymers), siliconecopolyols. Specific examples of non-ionic surfactants contemplated foruse include, but are by no means limited to, cetearyl alcohol,ceteareth-20, nonoxynol-9, C12-15 pareth-9, POE(4) lauryl ether,cocamide DEA, glycol distearate, glyceryl stearate, PEG-100 stearate,sorbitan stearate, PEG-8 laurate, polyglyceryl-10 trilaurate, laurylglucoside, octylphenoxy-polyethoxyethanol, PEG-4 laurate, polyglyceryldiisostearate, polysorbate-60, PEG-200 isostearyl palmitate, sorbitanmonooleate, polysorbate-80.

Non-limiting examples of preferred zwitterionic or amphotericsurfactants include betaines; sultaines; hydroxysultaines, amidobetaines, amidosulfo betaines; and combinations thereof. Specificexamples of amphoteric surfactants contemplated for use include, but areby no means limited to, cocoamidopropyl sultaine, cocoamidopropylhydroxyl sultaine, cocoamidopropylbetaine, coco dimethyl carboxymethylbetaine, lauryl dimethyl carboxymethyl betaine, lauryl dimethylalphacarboxyethyl betaine, cetyl dimethyl carboxymethyl betaine, cetyldimethyl betaine, lauryl (2-bishydroxy) carboxymethyl betaine, stearylbis-(2-hydroxyethyl) carboxymethyl betaine, oelyl dimethylgamma-carboxypropyl betaine, lauryl bis-(2-hydroxypropyl)alphacarboxymethyl betaine, coco dimethyl sulfopropyl betaine, stearyldimethyl sulfopropyl betaine, lauryl dimethyl sulfoethyl betaine, laurylbis(2-hydroxyethyl) sulfopropyl betaine, oleyl betaine, cocamidopropylbetaine.

In further accordance with the present invention, the isothiocyanatefunctional surfactant may optionally be incorporated into a formulationcomprising one or more solvents. Preferably, the solvent comprises ahydrocarbon and/or silicone oil that is generally non-hygroscopic and/orgenerally hydrophobic. Suitable examples, include, silicone basedsolvents and/or fluids, mineral oil, vegetable oils, squalene (i.e.,2,6,10,15,19,23-hexamethyltetracosane)—just to name a few.

The invention is further described by the following examples.

Example I Preparation of a mixture ofN_(α)-lauroyl-N_(ε)-isothiocyanato-L-Lysine withN_(α),N_(ε)-bis-lauroyl-L-lysine

A 1 liter beaker equipped with an overhead mechanical stainless steelpaddle stirrer was charged with 100 mL of 1 M NaOH (0.100 mol). Stirringwas begun and the beaker cooled to −5° C. to −10° C. using a salt/icebath. Next, 23.4 g (0.100 mol) of N_(ε)-benzylidene-L-lysine (preparedvia the method of Bezas, B and Zervas, L., JACS, 83, 1961, 719-722) wasadded. Immediately afterward and while keeping the solution cold, 140 mL(0.140 mol) of precooled (in a salt/ice bath) 1 M NaOH and 26.1 mL oflauroyl chloride was added in two equal portions over a period of 6minutes. The mixture was stirred for 10 more minutes at −5 to −10° C.,then the ice bath was removed and the reaction mixture allowed to stirfor another 1 hour while warming to room temperature. Next, the reactionmixture was cooled using a salt/ice bath and then sufficientconcentrated HCl was added to adjust the pH to 7.5-7.8. With the pH at7.8-7.8 and with continued cooling and stirring, 4.6 mL (60% ofstoichiometric, 0.068 mol) of thiophosgene was added drop-wise via anadditional funnel over the period of 1 hour. During this time,sufficient 1 M NaOH was added to maintain a pH range between 7.5-7.8.After the thiophosgene addition was complete, additional 1 M NaOH wasadded as necessary until the pH stabilized in 7.5-7.8 range. Next,sufficient 30% NaOH was added to adjust the pH to approximately 8.5.Next, 12 mL (0.051 mol) of lauroyl chloride was rapidly added, followedby sufficient 1 M NaOH to keep the pH in the range of 8.00-8.50. Next,sufficient concentrated HCl was added to adjust the pH to 1.5. Thereaction mixture was filtered via vacuum filtration, and the precipitatewashed with dilute HCl (pH=2). The product, a white moist solid, wasdried in vacuo while heating to 60° C. 45.19 g of white solid productwas recovered, a mixture of predominantlyN_(α)-lauroyl-N_(ε)-isothiocyanato-L-lysine andN_(α),N_(ε)-bis-lauroyl-L-lysine (determined via LC-MS analysis). Bothcompounds in this mixture can be simultaneously converted into anionic(carboxylate) surfactants via reaction with aqueous NaOH to yield aclear aqueous solution of the surfactants.

Example II Preparation of PureN_(α)-lauroyl-N_(ε)-isothiocyanato-L-Lysine Step 1: Preparation ofN_(α)-lauroyl-N_(ε)-carbobenzoxy-L-Lysine

60.0 g of N_(ε)-cbz-L-Lysine (cbz is carbobenzoxy) purchased fromAtomole Scientific Company, LTD was added to a three-liter beaker alongwith 1200 mL of RO water and the mixture was stirred. Next, 39 mL of 30%aqueous NaOH was added, resulting in dissolution of theN_(ε)-cbz-L-Lysine. The resulting solution was cooled in an ice bath andthen 52.5 mL of lauroyl chloride was added. The ice bath was removed 30minutes later, and stirring continued for an additional six hours, atwhich time 18 mL of concentrated hydrochloric acid was added. Thereaction mixture was then filtered via vacuum filtration, the whitesolid product washed with 1 M aqueous HCl, and then the solid productwas dried in vacuo while heated to approximately 85° C. 96.5 g of drywhite solid product was obtained. The product can be further purified bydissolving it in methanol, filtering off any insoluble precipitate, andremoving the methanol in vacuo to recover a white solid product (mp99.5-103.0° C.)

Step 2: Preparation of N_(α)-lauroyl-N_(ε)-ammonium chloride-L-Lysine

10.0 g of N_(α)-lauroyl-N_(ε)-carbobenzoxy-L-Lysine was weighed into aone liter Erlenmeyer flask equipped with a magnetic stir bar. 150 mL ofconcentrated hydrochloric acid was added and the solution was stirredand heated in an oil bath to 104° C., then allowed to cool with the oilbath back to room temperature. The solution was then cooled to 9° C. forapproximately four hours, during which time a large mass of whiteprecipitate formed. The reaction mixture was filtered in vacuo andrinsed with a small amount of cold 1 M HCl. The white solid reactionproduct was then dried in vacuo while being heated to 78° C., yielding7.89 g of white solid product (mp 191-193° C.).

Step 3: Preparation of N_(α)-lauroyl-N_(ε)-isothiocyanato-L-Lysine

0.46 mL of thiophosgene was added to 30 mL of dichloromethane in a 125mL Erlenmeyer flask equipped with a magnetic stir bar. To this solutionwas drop wise added over 15 minutes a solution consisting of 2.00 gN_(α)-lauroyl-N_(ε)-ammonium chloride-L-Lysine, 10 mL RO water, and 2.7mL 20% aqueous NaOH. Stirring was continued for an additional 30minutes, after which sufficient concentrated hydrochloric acid was addedto lower the pH to 1 as indicated by testing with pHydrion paper. Thereaction solution was then transferred into a separatory funnel and thebottom turbid dichloromethane layer was isolated and dried withanhydrous magnesium sulfate and gravity filtered. To the filtrate wasadded 50 mL of hexanes. The solution was then concentrated via removalof 34 mL of solvent via trap-to-trap distillation and then placed in a−19° C. freezer. A mass of white precipitate formed after a few hoursand was isolated via vacuum filtration and then dried in vacuo for 2hours. 1.130 g of a slightly off white solid powder product was obtained[mp 37.0-39.0° C.; IR (cm⁻¹), 3301sb, 2923s, 2852s, 2184m, 2099s, 1721s,1650s, 1531s, 1456m, 1416w, 1347m, 1216m, 1136w].

The oils and/or solvents employed hereinabove are provided for thepurposes of illustration, and are not to be construed as limiting theinvention in any way. As such, the oils may be liquid, solid, or gel,and may be synthetic or of natural origin and include but are notlimited to waxes, esters, lipids, fats, glycerides, cyclic silicones,linear silicones, crosslinked silicones, alkylsilicones, siliconecopolyols, alkylated silicone copolyols, and/or hydrocarbons, and/orethoxylated versions of all of these.

The foregoing description merely explains and illustrates the inventionand the invention is not limited thereto except insofar as the appendedclaims are so limited, as those skilled in the art who have thedisclosure before them will be able to make modifications withoutdeparting from the scope of the invention.

What is claimed and desired to be secured by Letters Patent of theUnited States is:
 1. A method for reducing macrophage migrationinhibitory factor cytokine or its biological activity, comprising thestep of: administering an isothiocyanate functional surfactant to apatient having a disease or condition wherein MIF cytokine or itsbiological activity is implicated in the disease or condition.
 2. Themethod according to claim 1, wherein the isothiocyanate functionalsurfactant comprises at least one isothiocyanate functional groupassociated with an aliphatic and/or aromatic carbon atom of theisothiocyanate functional surfactant.
 3. The method according to claim1, wherein the isothiocyanate functional surfactant comprises a lysinederivative, wherein the lysine derivative comprises an α-nitrogen and aε-nitrogen, and wherein an alkyl and/or alkanoyl substituent comprisingat least approximately 8 carbon atoms is associated with the α-nitrogen,and further wherein at least one isothiocyanate functional group isassociated with the ε-nitrogen.
 4. The method according to claim 1,wherein the isothiocyanate functional surfactant is represented by thefollowing chemical structure:

wherein the protonated form of the surfactant comprises a non-polarmoiety (NP) and a polar moiety (P), and wherein at least oneisothiocyanate functional group (NCS) is associated with the polarand/or non-polar moiety.
 5. The method according to claim 1, wherein theprotonated form of the isothiocyanate functional surfactant isrepresented by the following chemical structure:

wherein R₁ comprises an alkyl, cycloalkyl, polycycloalkyl,heterocycloalkyl, aryl, alkaryl, aralkyl, alkoxy, alkanoyl, aroyl,alkenyl, alkynyl and/or cyano group containing approximately 1 toapproximately 25 carbon atom(s), wherein the carbon atom(s) may be alinking group to, or part of, a halogen, a N, O, and/or S containingmoiety, and/or one or more functional groups comprising alcohols,esters, ammonium salts, phosphonium salts, and combinations thereof; alinkage to a dimer; a linkage to an oligomer; and/or a linkage to apolymer; wherein R₂ comprises NCS; and wherein R₃-R₅ are the same ordifferent and comprise H; OH; an alkyl, cycloalkyl, polycycloalkyl,heterocycloalkyl, aryl, alkaryl, aralkyl, alkoxy, alkanoyl, aroyl,alkenyl, alkynyl and/or cyano group containing approximately 1 toapproximately 25 carbon atom(s), wherein the carbon atom(s) may be alinking group to, or part of, a halogen, a N, O, and/or S containingmoiety, and/or one or more functional groups comprising alcohols,esters, ammonium salts, phosphonium salts, and combinations thereof; alinkage to a dimer; a linkage to an oligomer; and/or a linkage to apolymer with the proviso that at least one of R₃-R₅ comprise an alkyl,cycloalkyl, polycycloalkyl, heterocycloalkyl, aryl, alkaryl, aralkyl,alkoxy, alkanoyl, aroyl, alkenyl, alkynyl and/or cyano group containingapproximately 8 to approximately 25 carbon atom(s).
 6. The methodaccording to claim 1, wherein the protonated form of the isothiocyanatefunctional surfactant is represented by the following chemicalstructure:

wherein R₁ is selected from the group consisting of an alkyl groupcontaining 1 to 25 carbon atom(s); wherein R₂ is selected from the groupconsisting of NCS; and wherein R₃-R₅ are each independently selectedfrom the group consisting of H; OH; and an alkyl, and alkanoyl groupcontaining 1 to 25 carbon atom(s) with the proviso that at least one ofR₃-R₅ is selected from the group consisting of an alkyl, and alkanoyl,group containing 8 to 25 carbon atoms.
 7. The method according to claim1, wherein the protonated form of the isothiocyanate functionalsurfactant is represented by the following chemical structure:

wherein X comprises an integer ranging from approximately 1 toapproximately 25, and wherein Y comprises an integer ranging fromapproximately 6 to approximately
 25. 8. The method according to claim 1,wherein the protonated form of the isothiocyanate functional surfactantis represented by the following chemical structure:


9. The method according to claim 1, wherein the protonated form of theisothiocyanate functional surfactant is represented by at least one ofthe following chemical structures:


10. The method according to claim 1, wherein the isothiocyanatefunctional surfactant is represented by the following chemicalstructure:

wherein R₁ comprises an alkyl, cycloalkyl, polycycloalkyl,heterocycloalkyl, aryl, alkaryl, aralkyl, alkoxy, alkanoyl, aroyl,alkenyl, alkynyl and/or cyano group containing approximately 1 toapproximately 25 carbon atom(s), wherein the carbon atom(s) may be alinking group to, or part of, a halogen, a N, O, and/or S containingmoiety, and/or one or more functional groups comprising alcohols,esters, ammonium salts, phosphonium salts, and combinations thereof; alinkage to a dimer; a linkage to an oligomer; and/or a linkage to apolymer; wherein R₂ comprises NCS; wherein R₃-R₅ are the same ordifferent and comprise H; OH; an alkyl, cycloalkyl, polycycloalkyl,heterocycloalkyl, aryl, alkaryl, aralkyl, alkoxy, alkanoyl, aroyl,alkenyl, alkynyl and/or cyano group containing approximately 1 toapproximately 25 carbon atom(s), wherein the carbon atom(s) may be alinking group to, or part of, a halogen, a N, O, and/or S containingmoiety, and/or one or more functional groups comprising alcohols,esters, ammonium salts, phosphonium salts, and combinations thereof; alinkage to a dimer; a linkage to an oligomer; and/or a linkage to apolymer with the proviso that at least one of R₃-R₅ comprise an alkyl,cycloalkyl, polycycloalkyl, heterocycloalkyl, aryl, alkaryl, aralkyl,alkoxy, alkanoyl, aroyl, alkenyl, alkynyl and/or cyano group containingapproximately 8 to approximately 25 carbon atom(s), wherein X comprisesa counter cation such as, but not limited to, alkali metals, alkalineearth metals, transition metals, s-block metals, d-block metals, p-blockmetals, NZ₄ ⁺, wherein Z comprises, H, R₆, OR₆, and wherein R₆ comprisesan alkyl, cycloalkyl, polycycloalkyl, heterocycloalkyl, aryl, alkaryl,aralkyl, alkoxy, alkanoyl, aroyl, alkenyl, alkynyl and/or cyano groupcontaining approximately 1 to approximately 25 carbon atom(s), whereinthe carbon atom(s) may be a linking group to, or part of, a halogen, aN, O, and/or S containing moiety, and/or one or more functional groupscomprising alcohols, esters, ammonium salts, phosphonium salts, andcombinations thereof; a linkage to a dimer; a linkage to an oligomer;and/or a linkage to a polymer.
 11. The method according to claim 1,wherein the isothiocyanate functional surfactant is represented by thefollowing chemical structure:

wherein R₁ is selected from the group consisting of an alkyl groupcontaining 1 to 25 carbon atom(s); wherein R₂ is selected from the groupconsisting of NCS; and wherein R₃-R₅ are each independently selectedfrom the group consisting of H; OH; and an alkyl, and alkanoyl groupcontaining 1 to 25 carbon atom(s) with the proviso that at least one ofR₃-R₅ is selected from the group consisting of an alkyl, and alkanoyl,group containing 8 to 25 carbon atoms; and wherein X comprises a countercation.
 12. The method according to claim 1, wherein the isothiocyanatefunctional surfactant is administered to the patient at least one oforally, intravenously, intramuscularly, intrathecally, cutaneously,subcutaneously, transdermally, sublingually, buccally, rectally,vaginally, ocularly, otically, and nasally.
 13. The method according toclaim 12, wherein the amount of isothiocyanate functional surfactanttopically administered to the patient ranges from approximately 0.5nmol/cm2 to approximately 10 μmol/cm2.
 14. The method according to claim13, wherein the isothiocyanate functional surfactant is a topicalpreparation selected from the group consisting of ointment, cream,emulsion, lotion and gel.
 15. The method according to claim 14, whereinthe isothiocyanate functional surfactant further comprises one or morepharmaceutical, biological or molecular biological active agents. 16.The method according to claim 1, wherein the patient is a mammal. 17.The method according to claim 16, wherein the mammal is a human.